项目名称: TNF-α通过特定microRNA抑制喉黏膜间充质干细胞免疫调控作用的机制研究
项目编号: No.81470679
项目类型: 面上项目
立项/批准年度: 2015
项目学科: 医药、卫生
项目作者: 邓志宏
作者单位: 中国人民解放军第四军医大学
项目金额: 73万元
中文摘要: 慢性喉炎是发病率最高的呼吸道疾病之一,严重危害人体健康,其发病分子机制尚不清楚。本项目组前期研究发现间充质干细胞(MSCs)在喉黏膜组织的维持、修复及再生中发挥重要作用,并对局部炎症具有强大的调节功能,而TNF-α导致MSCs免疫调控功能下降,是炎症反应加剧的关键原因。我们发现慢性炎症组织来源MSCs的免疫调控能力受到不可逆的抑制,而miRNA介导的表观遗传调控对MSCs功能至关重要,据此推测炎症微环境通过表观遗传调控导致MSCs免疫功能异常。依此本项目拟对正常与炎症喉黏膜组织来源MSCs进行miRNA表达谱高通量筛选,明确关键miRNA,利用合成miRNA模拟物/抑制物、慢病毒过表达/干扰载体转染等手段研究其对于MSCs免疫调节功能的影响及靶基因,通过CHIP等技术明确TNF-α作用靶点及下游通路。从而探索miRNA表观遗传修饰对炎症MSCs的调控机制、探寻基于miRNA的炎症治疗策略。
中文关键词: 喉粘膜间充质干细胞;慢性炎症;免疫调控;表观遗传;微小RNA
英文摘要: Chronic laryngitis, which is the most common disease in respiratory tract, is harmful for local and systemic health. Our research group have identified a laryngeal mesenchymal stem cells (LMSCs), which play important role in laryngeal mucosa maintenance, repair and regeneration. Moreover, LMSCs are potent to regulate local inflammatory events. During chronic laryngitis, the immune-regulatory property of MSCs is inhibited by TNF-α, which might be a key factor to aggravate inflammation. But the molecular mechanism remains unknown. Our further research revealed that the inhibition of MSCs immune-regulatory property is inreversible. We also found that microRNA significantly regulated MSCs biological behavior through epigenetic pathway. Taken together, we hypothesized that TNF-α lead to dysfunction of MSCs through regulating miRNAs. In this study, we plan to perform miRNA micro-array screening between MSCs derived from normal and inflammatory mucosa; find out the key miRNAs; confirm the function of selected miRNAs and their targeting genes using strategies such as miRNAs mimics/inhibitor, and lentivirus overexpression/knockdown vector; determine the downstream regulatory pathway of TNF-α using CHIP and western blot analysis. The study will be helpful for us to understand undermining relationship between inflammation micro-environment and MSCs, to determine the mechanism of specific miRNAs regulation to MSCs, and to provide basic experimental evidence for novel strategy of inflammatory diseases.
英文关键词: laryngeal mesenchymal stem cell;chronic inflammation;inflammatory regulation;epigentics;microRNA